کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2035547 | 1072193 | 2014 | 14 صفحه PDF | دانلود رایگان |
• Simple and efficient brain-clearing reagents and image informatics were developed
• CUBIC is applicable to imaging of fluorescent proteins in adult brains and 3D-IHC
• The CUBIC method is scalable from a primate brain to subcellular structures
• CUBIC enables whole-brain imaging with single-cell resolution
SummarySystems-level identification and analysis of cellular circuits in the brain will require the development of whole-brain imaging with single-cell resolution. To this end, we performed comprehensive chemical screening to develop a whole-brain clearing and imaging method, termed CUBIC (clear, unobstructed brain imaging cocktails and computational analysis). CUBIC is a simple and efficient method involving the immersion of brain samples in chemical mixtures containing aminoalcohols, which enables rapid whole-brain imaging with single-photon excitation microscopy. CUBIC is applicable to multicolor imaging of fluorescent proteins or immunostained samples in adult brains and is scalable from a primate brain to subcellular structures. We also developed a whole-brain cell-nuclear counterstaining protocol and a computational image analysis pipeline that, together with CUBIC reagents, enable the visualization and quantification of neural activities induced by environmental stimulation. CUBIC enables time-course expression profiling of whole adult brains with single-cell resolution.
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Journal: - Volume 157, Issue 3, 24 April 2014, Pages 726–739